Abstract
In this paper, we propose an auto focusing method to find the best plane of focus amongst different depth micro objects. By achieving this, the ultimate goal of the project will be autofocusing on motile micro-organisms to check their quality, e.g., semen motility. Firstly, the auto focusing system cycles through the rough focal range of the micro objects, which represents the cell culture by moving in steps of 2 \(\upmu \hbox {m}\) towards best focus, passing it, and completing the scanning process. Simultaneously, at each step, an image of the sample is collected and transformed by use of a Prewitt edge detection operator in order to ascertain focal quality. Secondly, the system moves to the best focus position that is calculated via Prewitt edge detection approach. Thirdly, to begin actively monitoring the focal plane, the system makes minute adjustments to keep the target objects in-focus. This accounts for any changes in the culture, such as cells moving in and out of the focal window or up and down within the semen fluid. The Prewitt operator is tested for focusing performance on multi-microobjects at different focal planes and is compared with other edge detection gradient functions to confirm superiority.





















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Baraniuk, R.G.: Laplacian Edge Detection. https://www.owlnet.rice.edu/~elec539/Projects97/morphjrks/laplacian.html (1995)
Bhaktia, T.L., Susantoa, A., Santosaa, P.I., Tri, D.: Low temperature sperm selection method to support Bovine breeding industry. J. Teknol. 78(5–9), 125–131 (2016)
Broekhuijse, M.L.W.J., Otari, E., Feitsma, H., Gadella, B.M.: Additional value of computer assisted semen analysis (CASA) compared to conventional motility assessments in pig artificial insemination. Theriogenology 76(8), 1473–1486 (2011)
Chen, C.H., Feng, T.L.: Fast 3D shape recovery of a rough mechanical component from real time passive autofocus system. Int. J. Adv. Manuf. Technol. 34(9–10), 944–957 (2007)
Cho, J.H., Cho, S.B.: An architecture for real-time hardware cosimulation of edge detection in image processing using Prewitt edge operator. In: International Conference on Electronics, Information and Communications (ICEIC), pp. 1–2 (2014)
David, I., Kohnke, P., Fehrenbach, J., Simoes, A.L., Debreuve, E., Descombes, X., Druart, X.: New objective measurements of semen wave motion are associated with fertility in sheep. Reprod. Fertil. Dev. 30(6), 889–896 (2018)
Dott, H.M., Foster, G.C.A.: The estimation of sperm motility in semen, on a membrane slide, by measuring the area change frequency with an image analysing computer. J. Reprod. Fertil. 55(1), 161–166 (1979)
Elmabrouk, A., Aggoun, A.: Edge detection using local histogram analysis. Electron. Lett. 34(12), 1216–1217 (1998)
Giwercman, A., Richthoff, J., Hjllund, H., Bonde, J.P., Jepson, K., Frohm, B., Spano, M.: Correlation between sperm motility and sperm chromatin structure assay parameters. Fertil. Steril. 80(6), 1404–1412 (2003)
Groen, F.C., Young, I.T., Ligthart, G.: A comparison of different focus functions for use in autofocus algorithms. Cytometry J. Int. Soc. Anal. Cytol. 6(2), 81–91 (1985)
Hsu, W.Y., Lee, C.S., Chen, P.J., Chen, N.T., Chen, F.Z., Yu, Z.R., Hwang, C.H.: Development of the fast astigmatic auto-focus microscope system. Meas. Sci. Technol. 20(4), 045902 (2009)
Innings, F., Trägårdh, C.: Visualization of the drop deformation and break-up process in a high pressure homogenizer. Chem. Eng. Technol. Ind. Chem. Plant Equip. Process Eng. Biotechnol. 28(8), 882–891 (2005)
Jacobs, D.: Image gradients. Class Notes CMSC 2005, 426 (2005)
James, Tomlinson, M., Pooley, K., Simpson, T., Newton, T., Hopkisson, J., Jayaprakasan, K., Jayaprakasan, R., Naeem, A., Pridmore, T.: Validation of a novel computer-assisted sperm analysis (CASA) system using multitarget-tracking algorithms. Fertil. Steril. 93(6), 1911–1920 (2010)
Jiwen, C., Jiubin, T., Lei, A.: Automatic focusing method based on fuzzy control and image processing technique. Opto-Electronic Engineering, pp. 12–16 (2005)
Kanopoulos, N., Vasanthavada, N., Baker, R.L.: Design of an image edge detection filter using the Sobel operator. IEEE J. Solid State Circ. 23, 2 (1988)
Liu, C.S., Hu, P.H., Wang, Y.H., Ke, S.S., Lin, Y.C., Chang, Y.H., Horng, J.B.: Novel fast laser-based auto-focusing microscope. IEEE Sens. 1, 481–485 (2010)
Manjula, V.S.: Image edge detection and segmentation by using histogram thresholding method. J. Eng. Res. Appl. 7(8), 10–16 (2017)
Miaofen, Z., Wanqiang, W., Guojin, C., Yongning, L.: Image focusing system based on FPGA, in Computer Communication Control and Automation (3CA), 1, 415-418 (2010)
Nayar, S.K., Nakagawa, Y.: Shape from focus. IEEE Trans. Pattern Anal. Mach. Intell. 16(8), 824–831 (1992)
Nguyen, C.N., Ohara, K., Takubo, T., Mae, Y., Arai, T.: Highspeed autofocusing of multisized microobjects. In: IEEE International Conference on Automation Science and Engineering (CASE), pp. 34–39 (2012)
Obara, T., Igarashi, Y., Hashimoto, K.: Fast and adaptive autofocusing algorithm for microscopic cell observation. In: IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS), pp. 7–12 (2011)
Ohara, K., Kojima, M., Takagi, S., Horade, M., Mae, Y., Arai, T.: Development of the 3D measurement system in real-time for micromanipulation. In: IEEE International Conference on Mechatronics and Automation (ICMA), pp. 2022–2027 (2017)
Ohba, K., Ortega, J.C.P., Tanie, K., Tsuji, M., Yamada, S.: Microscopic vision system with all-in-focus and depth images. Mach. Vis. Appl. 15(2), 55–62 (2003)
Palacin, I., Vicente-Fiel, S., Santolaria, P., Yaniz, J.L.: Standardization of CASA sperm motility assessment in the ram. Small Rumin. Res. 112(1–3), 128–135 (2013)
Priyanto, H., Awaludin, I., Sukmawati, E., Utami, D., Nuriyadi, M.: Bull sperm motility measurement using improved matching-based algorithm and ellipse detection. In: Informatics and Computing (ICIC), International Conference, pp. 55–60 (2016)
Russel, M.J., Douglas, T.S.: Evaluation of autofocus algorithms for tuberculosis microscopy. In: Engineering in Medicine and Biology Society (EMBS), pp. 3489–3492 (2007)
Salisbury, G.W., Fuller, H.K., Willett, E.L.: Preservation of bovine spermatozoa in yolk-citrate diluent and field results from its use. J. Dairy Sci. 24(11), 905–910 (1941)
Shapiro, H., Shapiro, L.: Computer and Robot Vision, vol. 1, pp. 345–351. Addison-Wesley Publishing Company, Pearson (1992)
Song, Y., Li, M.: Research on SUSAN based auto-focusing algorithm for optical microscope application. In: IEEE International Conference on Mechatronics and Automation, pp. 1237–1241 (2006)
Taute, K.M., Gude, S., Tans, S.J., Shimizu, T.S.: High-throughput 3D tracking of bacteria on a standard phase contrast microscope. Nat. Commun. 6, 8776 (2015)
Wang, F.Y., Wang, R., Zhang, G.J.: Auto-focusing system based on DSP [J]. Opto-Electron. Eng. 8, 030 (2007)
Watanabe, M., Nayar, S.K.: Minimal operator set for passive depth from defocus in proceedings. IEEE Computer Society Conference on Computer Vision and Pattern Recognition CVPR96, pp. 431–438 (1996)
World Health Organisation: WHO Laboratory Manual for the Examination of Human Semen and Sperm-Cervical Mucus Interaction. Cambridge University Press, Cambridge (1999)
Zeder, M., Pernthaler, J.: Multispot liveimage autofocusing for highthroughput microscopy of fluorescently stained bacteria. Cytometry Part A 75(9), 781–788 (2009)
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Research supported by Massey University Research Fund (MURF) 2017.
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Lofroth, M., Avci, E. Auto-focusing approach on multiple micro objects using the prewitt operator. Int J Intell Robot Appl 2, 413–424 (2018). https://doi.org/10.1007/s41315-018-0070-x
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DOI: https://doi.org/10.1007/s41315-018-0070-x